Fig. 3: SAM guide delivery approaches.

a AAV8^3aTtr (n = 3) and AAV8^GFP (n = 2) were delivered to R26^SAM/+ mice. ELISA was employed to determine protein levels along with untreated R26^SAM/+ (n = 2). Values were plotted as the mean per group +/− SD. Monthly p-values can be found in Supplementary Table 3. Two independent studies were conducted with similar results. b AAV8^4aTsx was delivered to R26^SAM/+ mice (n = 3) to evaluate the tissue-specific induction of Tsx. Expression values relative to R26^SAM/+ were plotted as the mean per group +/− SD. Liver: p = 0.001026; Spleen: p = 0.01138. c Protein induction by arrayed guide viral delivery to R26^SAM/SAM mice was compared to single guide viral delivery (n = 3 for controls, n = 6 for treated). Serum levels were plotted as the mean per group +/− SD. p-values of each array comparison can be found in Supplementary Table 3. d AAV8^4aTsx comprises two guides targeting the 300 bp directly upstream of the TSS and another two guides targeting an upstream enhancer region (750 bp upstream). qPCR was completed on livers harvested from R26^SAM/SAM treated with either AAV delivered guide array (4aTsx) or individual guides from the array (Tsx gA1-4) (n = 5). Expression values relative to R26^SAM/+ were plotted as the mean per group +/− SD. p-values of each array comparison can be found in Supplemental Table 3. e LNP particles were formulated with synthetic Ttr gA3 SAM guides and introduced to R26^SAM/SAM mice (n = 3). Protein expression levels were determined by ELISA with weekly bleeds. Daily p-values can be found in Supplementary Table 3. All values are plotted as mean +/− SD. Two independent replicates were conducted with similar results. f LNP particles were formulated with 0.5 mpk of synthetic Ttr gA2 SAM guide and introduced to R26^SAM/SAM mice (n = 5) at zero weeks and/or 4 weeks. Protein expression levels were determined by ELISA with weekly bleeds. Daily p-values can be found in Supplementary Table 3. All values are plotted as mean +/− SD. g Overexpression by HDD delivery of Ttr cDNA or a plasmid expressing arrayed SAM guides was compared to AAV8^3aTtr in R26^SAM/SAM mice (n = 5, per group). TTR levels were determined by ELISA and plotted as mean per group +/− SD. Untreated vs Array week 1: p = 0.000001; Untreated vs. Array week 2: p = 0.000001; Untreated vs HDD^3aTtr week 1: p = 0.003585; Untreated vs. HDD^Ttr plasmid week 1: p = 0.002222. Two independent replicates were conducted with similar results. h A guide array expressing two activating guides each to Ngn3 and Btc was delivered to R26^SAM/SAM mice (n = 2). Five tissues were collected and expression relative to R26^SAM/SAM was plotted as mean per group +/− SD. Statistics: Asterisks (*) indicates significance, and the number of asterisks indicates the number of 0s after the decimal point. One-tailed, unpaired Student’s t-test for (a–j).